Skip to main content
. Author manuscript; available in PMC: 2017 Nov 27.
Published in final edited form as: Int J Biochem Cell Biol. 2016 Aug 30;79:139–150. doi: 10.1016/j.biocel.2016.08.023

Fig. 6. Inhibition of kinase activity and knockdown of VEGF receptors decrease leptin induction of Notch expression in endothelial cells.

Fig. 6

A. Leptin induction of Notch receptors in HUVEC requires VEGFR2 kinase activity. Western blot representative results from the reduction of leptin-induced Notch receptors via VEGFR kinase inhibition. HUVEC were treated for 24 h with leptin (1.2 nM), VEGFR-2 inhibitor SU5416 (5 µmol/l) and positive control VEGF (25 ng/ml). B. Leptin induction of Notch proteins in HUVEC requires VEGFR-2 gene activity. Represented here are western blot results from the reduction of leptin-induced Notch via VEGFR-2 SiRNA knockdown. Results show the effects of VEGFR-2 SiRNA (1 µg) after 6 h on leptin-induced expression of Notch receptors and ligands in HUVEC. C. Leptin induction of Notch proteins in HUVEC is reduced by knockdown of VEGFR-1 and VEGFR-2 gene activities. Western blot representative results from the reduction of leptin-induced Notch via VEGFR-1 and VEGFR-2 SiRNA knockdown. D. Leptin induction of Notch mRNA in HUVEC is reduced by knockdown of VEGFR-1 and VEGFR-2 gene activities. Quantitative results from real-time PCR of Notch, VEGFR-1 and VEGFR-2 mRNA expression in HUVEC treated with leptin and VEGFR-1 and VEGFR-2 SiRNA. Beta-actin or GAPDH were used as loading controls. Relative protein expression was calculated as percentage to basal. Histograms show densitometric analysis of Notch protein expression using NIH image J software. RNA expression was calculated by normalizing values to GAPDH mRNA. Relative mRNA expression was calculated to basal. Data is presented as an average ± s.d. from three independent experiments. Su: SU5416; SiControl: siRNA control. a: p<0.05 when compared to basal. b: p<0.05 when compared to endothelial cells treated with leptin.